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MicroRNA-1 promotes cartilage matrix synthesis and regulates chondrocyte differentiation via post-transcriptional suppression of Ihh expression

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Indian hedgehog signaling molecule (Ihh) is known to play critical roles in chondrogenesis and cartilage development. However, it remains largely unknown how Ihh is regulated during the process. Previous studies… Click to show full abstract

Indian hedgehog signaling molecule (Ihh) is known to play critical roles in chondrogenesis and cartilage development. However, it remains largely unknown how Ihh is regulated during the process. Previous studies suggest that Ihh plays an important regulatory role in the growth and development of articular cartilage, but whether it is regulated by miRNAs is unclear. The present study aimed to investigate the effects of miR-1 on chondrocyte differentiation and matrix synthesis, and to determine whether miR-1 can regulate the Ihh signaling pathway. In the present study, the expression level of miR-1 was altered via transfection of the miR-1 mimic or inhibitor in mouse thorax chondrocytes, and the impact on chondrocyte phenotypes and Ihh expression was examined. Overexpression of miR-1 promoted the expression of the matrix synthesis-associated molecules collagen (Col)-II and aggrecan, two key components in cartilage matrix. Conversely, overexpression of miR-1 significantly downregulated the expression of chondrocyte differentiation markers Col-X and matrix metallopeptidase 13. Moreover, overexpression of miR-1 dose-dependently inhibited endogenous Ihh expression, and an association was observed between miR-1 and Ihh expression. The 3′ untranslated region (UTR) of Ihh from various species contains two miR-1 binding sites. Luciferase reporter assays indicated that miR-1 post-transcriptionally suppressed Ihh expression, which was dependent on the binding of miR-1 to one of the two putative binding sites of the Ihh 3′UTR. Furthermore, via inhibition of Ihh expression, miR-1 decreased the expression of molecules downstream of Ihh in the Hedgehog signaling pathway in mouse thorax chondrocytes. This study provided new insight into the molecular mechanisms of miR-1 in regulating chondrocyte phenotypes via targeting the Ihh pathway.

Keywords: ihh; chondrocyte differentiation; expression; ihh expression; matrix synthesis

Journal Title: Molecular Medicine Reports
Year Published: 2020

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